miR-105 通过靶向调控 SOX9 增强 hADSCs 的成骨分化。
MiR-105 enhances osteogenic differentiation of hADSCs via the targeted regulation of SOX9.
机构信息
Department of Orthopedic, Qinghai Provincial People's Hospital, Xining, Qinghai, China.
Department of Endocrinology, Qinghai University Affiliated Hospital, Xining, Qinghai, China.
出版信息
Tissue Cell. 2021 Oct;72:101540. doi: 10.1016/j.tice.2021.101540. Epub 2021 Apr 2.
OBJECTIVE
To investigate whether miR-105 can regulate the osteogenic differentiation of human adipose-derived mesenchymal stem cells (hADSCs) by targeting SOX9.
METHODS
The hADSCs were grouped for subsequent transfection and induction of osteogenic differentiation as follows: control, miR-NC, miR-105 mimics, miR-105 inhibitors, SOX9, SOX9 siRNA, miR-105 mimics + SOX9 and miR-105 inhibitors + SOX9 siRNA groups. Next, hADSCs were stained for alkaline phosphatase (ALP), and Alizarin Red S staining (ARS) was performed. Osteogenic differentiation-related genes and miR-105 expression were assessed by qRT-PCR, while SOX9 protein expression was determined by Western blotting.
RESULTS
MiR-105 expression was increased and SOX9 protein expression was decreased during the osteogenic differentiation of hADSCs. A dual-luciferase reporter assay confirmed SOX9 to be a target gene of miR-105. Compared with the control group, the miR-105 mimics and SOX9 siRNA groups had elevated BMP2, OPN, OCN, BSP, Osx and Runx2 mRNA expression with reduced SOX9 expression, as well as increased ARS intensity and ALP activity. After transfection of miR-105 inhibitors/SOX9 into hADSCs, the results were the opposite. Overexpressing SOX9 reversed the effect of miR-105 in promoting the osteogenic differentiation of hADSCs.
CONCLUSION
MiR-105 could target SOX9 to improve the expression of osteogenic differentiation genes and thus enhance the osteogenic differentiation of hADSCs.
目的
探讨 miR-105 是否可以通过靶向 SOX9 来调节人脂肪间充质干细胞（hADSCs）的成骨分化。
方法
将 hADSCs 分组进行转染和诱导成骨分化，具体分组如下：对照组、miR-NC 组、miR-105 模拟物组、miR-105 抑制剂组、SOX9 组、SOX9 siRNA 组、miR-105 模拟物+SOX9 组和 miR-105 抑制剂+SOX9 siRNA 组。然后对碱性磷酸酶（ALP）进行染色，并用茜素红 S 染色（ARS）进行检测。通过 qRT-PCR 评估成骨分化相关基因和 miR-105 的表达，通过 Western blot 检测 SOX9 蛋白表达。
结果
在 hADSCs 的成骨分化过程中，miR-105 的表达增加，SOX9 蛋白的表达减少。双荧光素酶报告基因检测证实 SOX9 是 miR-105 的靶基因。与对照组相比，miR-105 模拟物和 SOX9 siRNA 组的 BMP2、OPN、OCN、BSP、Osx 和 Runx2 mRNA 表达上调，SOX9 表达下调，ARS 强度和 ALP 活性增加。将 miR-105 抑制剂/ SOX9 转染 hADSCs 后，结果相反。过表达 SOX9 逆转了 miR-105 促进 hADSCs 成骨分化的作用。
结论
miR-105 可以靶向 SOX9 提高成骨分化基因的表达，从而增强 hADSCs 的成骨分化。
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